1. ProductProfile
EpiTect® Methyl II PCR Array System
For pathway- or disease-focused profiling of regional DNA methylation, without bisulfite
conversion, using MethylScreen™ Technology
The EpiTect Methyl II PCR Array System is an innovative technology enabling fast and accurate detection of CpG island DNA
methylation of individual genes, as well as disease- and pathway-focused gene panels. This technology does not require
bisulfite conversion. With a simple restriction enzyme digestion followed by real-time PCR, the DNA methylation levels of 22
or 94 different genes can be analyzed simultaneously using DNA Methylation PCR Arrays. The simplicity of the procedure
makes this technology highly suited for profiling DNA methylation and biomarker characterization. The EpiTect Methyl II PCR
Array System uses the MethylScreen Technology provided under license from Orion Genomics, LLC.
EpiTect Methyl II PCR System provides:
Workflow of EpiTect Methyl II PCR Array System
„„ Combined reliability of controlled restriction digestion with qPCR detection
Isolate DNA
sample
„„ Genomewide, ready-to-use, predesigned primer assays
„„ Profiling of multiple targets simultaneously in a single qPCR run
Incubate with
restriction
enzymes
„„ Free Web-based and automated data analysis tools
„„ Controls to monitor enzymatic activities of restriction digestion enzymes
No
Methylation- Methylationenzyme
sensitive
dependent
(Mo)
enzyme
enzyme
(Ms)
(Md)
MSRE
and
MDRE
(Msd)
Sensitivity of EpiTect Methyl II Assays
Methylated
Unmethylated
SEC
Mo
DEC
100
Ms
90
80
Add to array
Md
70
Msd
60
50
Real-time PCR
Methylated
Unmethylated
40
30
100
% Total input DNA
20
10
0
0.0
% SKBR3 genomic DNA
Analyze data
40
20
MGMT
ESR1
HIC1
GSTP1
CDKN2A
Figure 1. EpiTect Methyl II PCR assay detects methylation in heterogeneous samples. The analytical
sensitivity of the EpiTect Methyl II PCR Assay was tested using a serial dilution of SKBR3 cell line
genomic DNA and peripheral blood leukocyte genomic DNA. Using Human HIC1 DNA EpiTect
Methyl II PCR Primers, the percentage of methylated HIC1 relative to the total amount of input
DNA was detectable in each mixture even down to only 6% of the total DNA sample. The HIC1
gene promoter is methylated in cancer cells and unmethylated in normal cells.
CDKN2A
0
CDH1
6.3
CDH13
12.5
CDKN1C
33.3
CCND2
50.0
BRCA1
75.0
60
CCNA1
100.0
80
ADAM23
% Total input DNA
Mix remaining
DNA with SYBR®
Green qPCR
Mastermix
Figure 2. Schematic view of the EpiTect Methyl II PCR Array
System procedure. The system relies on the differential cleavage
of target sequences by two different restriction endonucleases.
qPCR allows the analysis of the methylation status of up to 94
targets simultaneously. SEC and DEC are control assays for
monitoring enzymatic activities of restriction digestion enzymes.
Sample & Assay Technologies
